This project will define precisely the identity and positions of all transcripts within the genome of the human fungal pathogen Candida albicans, by using ultra high throughput RNA sequencing, in conjunction with a rationally chosen diverse set of experimental conditions. C. albicans is an increasingly important human pathogen, and, due to an expanding patient population and to increasing resistance to existing antifungal drugs, it is vital that new treatments are developed. It is likely that hundreds of new transcripts will be identified using our approach, and many of these may be specific to C. albicans or closely related species;these novel pathogen-specific genes may prove to be good therapeutic targets. In addition, we will validate all new transcripts by RT-PCR, and identify which of the new transcripts are likely to encode proteins. Finally, we will perform preliminary functional characterization of the novel transcripts, by using microarrays to determine their relative abundance under the various used experimental conditions, and by identifying with which genes of known function they are co-regulated. This proposal is significant both because it directly advances our understanding of the biology of this medically important human fungal pathogen, and because similar studies from other organisms strongly suggest that the proposed experiments will substantially refine the primary annotation of the C. albicans genome. We thus gain insight into C. albicans biology and also benefit the field of fungal genomics more widely, with the long-term goal of having a significant impact into the way in which fungal infections are treated in human patients. With a chromosome-level assembly of the C. albicans genome, the existence of high throughput sequencing technologies, and a public Candida genomics database, the groundwork is in place for both successful completion of this project and dissemination of the data to the fungal pathogenesis research community to significantly impact future C. albicans research, and, ultimately the treatment of Candida disease. PUBLIC HEALTH RELEVANCE: Candida albicans is an increasingly important human pathogen, and the development of new treatments is vital, especially given the recent rise in drug resistance among clinical isolates of C. albicans. This work seeks to identify novel genes within C. albicans;it is likely that hundreds of new genes will be identified, and of those, many may prove to be good therapeutic targets. Thus, this work has the potential for a large positive future impact on human health.